FISH of urothelial cells Vittorio Grazioli, MD CEO – IMPACT LAB

The real cure of cancer is early diagnosis Cancer is the uncontrolled growth and spread of cells which often invade surrounding tissue and can metastasize to distant sites. Despite ever-increasing knowledge concerning the prevention and treatment of cancer, the number of new cases continues to grow each year. Currently, effective treatment of cancer relies on its early and accurate diagnosis.

Dysplasia is the frontline of the war against cancer Since 1932 (thanks to Dr. Broder) we have considered in situ carcinoma as cancer and, more recently, to recognize dysplasia as a pre-cancerous lesion. Dysplasia is a sort of grey zone, in which both regression and progression are possible, depending on factors that may represent a selective advantage for cancer cells but which cannot be identified by means of conventional methods.

From chromosomal instability to chromosomal chaos

From chromosomal instability to chromosomal chaos In the squamous epithelium (i.e. cervical, oral, anal) but also in the urothelial cells, the gain of chromosome 3 (whose 3q26 locus maps for telomerase) represents a selective advantage leading to cell immortalization and therefore to the progressive chromosomal chaos typical of cancer cells.

From chromosomal instability to chromosomal chaos NORMAL CELL TUMOR CELL

FISH Results - Negative Specimen Cytology review of a normal urine specimen. Normal specimen analyzed by FISH.

FISH Results - Positive Specimen Cytology review of an abnormal urine specimen. Same specimen analyzed by FISH.

WHAT DO WE NEED TO FISH Target DNA Probe Fluorescence microscope Cytogeneticist 9

CYTOPATHOLOGIC CLASSIFICATION OF URINE CYTOLOGY Koss Classification Negative Mild atypia Moderate atypia, suspect low-grade tumor Severe atypia, suspect high grade tumor Positive Rosenthal&Raab Classification Negative Benign cellular changes Atypical,indeterminatefor neoplasia Low grade neoplasia High grade neoplasia Classification used at IMPACT LAB Negative Mild/moderate atypia Reactive Cannot exclude low grade tumor Atypia, suspect for urothelial neoplasm Positive

DIAGNOSTIC CATEGORIES PREFERRED BY UROLOGISTS NEGATIV FOR HGUC SUSPICIOUS FOR HGUC POSITIVE FOR HGUC

SHOULD WE ELIMINATE THE “ATYPICAL” CATEGORY ? APPROX 10-20 % OF URINE ARE CLASSIFIED AS ATYPICAL THEREFORE, UP TO 80 % OF ABNORMAL URINE ARE CLASSIFIED AS ATYPICAL SIGNIFICANT PROPORTION OF MALIGNANT CASES WOULD BE MISSED IF “ATYPIA” WAS ELIMINATED

FISH: PRE-ANALYTICAL ISSUES PATIENT SELECTION INFLAMMATION SPECIMEN COLLECTION

FISH: REPORT READING AT LEAST 50 (IDEALLY 100) ATIPYCAL CELLS EVALUATED RESULT MUST BE QUANTITATIVE > 5% POSITIVITY = > 60% RISK